Metabolic tagging and targeting of red blood cells

Abstract

Modified red blood cells (RBCs) including one or more surface proteins or lipids covalently linked to a cargo by click chemistry are provided. In some examples, the cargo is a cancer therapeutic agent, an autoimmune antigen, or an antigen of a bacterial or viral agent. Also provided are methods of treating a subject with a disease or disorder with the modified RBCs or methods of treating a subject with a disease or disorder or performing imaging analysis of a subject, including administering to the subject a composition including an azido-modified sugar moiety, thereby generating red blood cells comprising one or more azido-labeled surface proteins and administering to the subject a composition including a cargo for treating or inhibiting the disease or disorder, wherein the cargo is capable of covalently binding to the one or more azido-labeled surface proteins or lipids.

We claim: 
     
         1 . A method for treating or inhibiting a disease, comprising:
 administering to a subject a composition comprising an azido-modified sugar moiety, thereby generating red blood cells comprising one or more azido-labeled surface proteins; and   administering to the subject a composition comprising a cargo for treating or inhibiting the disease, wherein the cargo is capable of covalently binding to the one or more azido-labeled surface proteins or lipids.   
     
     
         2 . The method of  claim 1 , wherein the composition comprising the cargo for treating or inhibiting the disease is administered to the subject after the composition comprising the azido-modified sugar moiety. 
     
     
         3 . The method of  claim 2 , wherein the composition comprising the cargo for treating or inhibiting the disease is administered to the subject at least about 3 days after the composition comprising the azido-modified sugar moiety. 
     
     
         4 . The method of  claim 1 , wherein the composition comprising an azido-modified sugar moiety, the composition comprising a cargo for treating or inhibiting the disease, or both is administered to the subject intravenously. 
     
     
         5 . The method of  claim 1 , wherein the azido-labeled sugar moiety is tetra-acetylated N-azidoacetyl-D-mannosamine (Ac 4 ManNAz), N-azidoacetyl-D-mannosamine (ManNAz), tetra-acetylated N-azidoacetyl-D-galactosamine (Ac 4 GalNAz), N-azidoacetyl-D-galactosamine (GalNAz), tetra-acetylated N-azidoacetyl-D-glucosamine (Ac 4 GlcNAz), N-azidoacetyl-D-glucosamine (GlcNAz), or N-Acetyl-9-azido-9-deoxy-neuraminic acid (9-AzNeu5Ac). 
     
     
         6 . The method of  claim 1 , wherein the cargo capable of covalently binding to the one or more azido-labeled surface proteins or lipids is capable of covalent binding by click chemistry. 
     
     
         7 . The method of  claim 6 , wherein the click chemistry is azide-alkyne click chemistry. 
     
     
         8 . The method of  claim 1 , wherein the cargo capable of covalently binding to the one or more azido-labeled surface proteins or lipids comprises dibenzocyclooctyne. 
     
     
         9 . The method of  claim 1 , wherein:
 the cargo is a cancer therapeutic compound and the disease is cancer; or   the cargo is insulin and the disease is diabetes; or   the cargo is an antigen and the disease is an infection; or   the cargo is an autoantigen and the disease is an autoimmune disorder; or   the cargo is an antibiotic and the disease is a bacterial infection; or   the cargo is an antibiotic and the disease is a viral infection.   
     
     
         10 . A method of in vivo imaging, comprising:
 administering to a subject a composition comprising an azido-modified sugar moiety, thereby generating red blood cells comprising one or more azido-labeled surface proteins;   administering to the subject a composition comprising an imaging agent, wherein the imaging agent is capable of covalently binding to the one or more azido-labeled surface proteins or lipids; and   performing an imaging analysis of the subject one or more times.   
     
     
         11 . The method of  claim 10 , wherein the imaging agent is a contrast agent, a fluorescent dye, or a bioluminescent substrate. 
     
     
         12 . The method of  claim 10 , wherein the imaging analysis is magnetic resonance imaging. 
     
     
         13 . A method for treating or inhibiting a disease, comprising:
 contacting a preparation of red blood cells obtained from a subject with the disease or disorder with a composition comprising an azido-modified sugar for a sufficient period of time for the red blood cells to express one or more azido-labeled surface proteins or lipids;   contacting the red blood cells expressing one or more azido-labeled surface proteins or lipids with a composition comprising a cargo for treating or inhibiting the disease, wherein the cargo is capable of covalently binding to the one or more azido-labeled surface proteins or lipids, thereby producing a population of modified red blood cells; and   administering the population of modified red blood cells to the subject.   
     
     
         14 . The method of  claim 13 , wherein the azido-labeled sugar moiety is tetra-acetylated N-azidoacetyl-D-mannosamine (Ac 4 ManNAz), N-azidoacetyl-D-mannosamine (ManNAz), tetra-acetylated N-azidoacetyl-D-galactosamine (Ac 4 GalNAz), N-azidoacetyl-D-galactosamine (GalNAz), tetra-acetylated N-azidoacetyl-D-glucosamine (Ac 4 GlcNAz), N-azidoacetyl-D-glucosamine (GlcNAz), or N-Acetyl-9-azido-9-deoxy-neuraminic acid (9-AzNeu5Ac). 
     
     
         15 . The method of  claim 13  wherein the population of red blood cells from the subject is contacted with the azido-labeled sugar moiety for about 1-120 hours. 
     
     
         16 . The method of  claim 13 , wherein the cargo capable of covalently binding to the one or more azido-labeled surface proteins or lipids is capable of covalent binding by click chemistry. 
     
     
         17 . The method of  claim 16 , wherein the click chemistry is azide-alkyne click chemistry. 
     
     
         18 . The method of  claim 13 , wherein the cargo capable of covalently binding to the one or more azido-labeled surface proteins or lipids comprises dibenzocyclooctyne. 
     
     
         19 . A modified red blood cell comprising one or more surface proteins or lipids covalently linked to a cargo by click chemistry. 
     
     
         20 . The modified red blood cell of  claim 19 , wherein the click chemistry is azide-alkyne click chemistry, tetrazine-norbornene click chemistry, tetrazine-cyclooctene click chemistry, or maleimide-thiol click chemistry. 
     
     
         21 . The modified red blood cell of  claim 19 , wherein the cargo is a cancer therapeutic compound, insulin, an antigen, an autoantigen, an antibiotic, an antiviral, an antibody, or an imaging agent. 
     
     
         22 . The modified red blood cell of  claim 19 , wherein the click chemistry comprises an azido-labeled sugar moiety. 
     
     
         23 . The modified red blood cell of  claim 22 , wherein the azido-labeled sugar moiety is tetra-acetylated N-azidoacetyl-D-mannosamine (Ac 4 ManNAz), N-azidoacetyl-D-mannosamine (ManNAz), tetra-acetylated N-azidoacetyl-D-galactosamine (Ac 4 GalNAz), N-azidoacetyl-D-galactosamine (GalNAz), tetra-acetylated N-azidoacetyl-D-glucosamine (Ac 4 GlcNAz), N-azidoacetyl-D-glucosamine (GlcNAz), or N-Acetyl-9-azido-9-deoxy-neuraminic acid (9-AzNeu5Ac). 
     
     
         24 . A composition comprising the modified red blood cell of  claim 19  and a pharmaceutically acceptable carrier.